Plin transgenic mice - Atherosclerosis
I. Purpose
Cells in humans synthesize and store intracellular lipid droplets (LDs). LDs have multiple biological roles, e.g. as efficient reservoirs of energy and substrates for membrane and steroid hormone synthesis. Perilipin1-5 proteins are abundant on the LD surface and are important for regulation of LD degradation. Development of Atherosclerosis starts with accumulation of esterfied cholesterol, free cholesterol and cholesterol crystals in the arteria intima. The purpose of this project is to understand the biological role of LDs and Plin proteins in development of atherosclerosis using mice with deleted Plin1-5 genes.
According to §10, the purpose is: a) Basal research.
II. Expected distress
Mice with single mutation of Plin genes give offspring with normal Mendalian distribution with no observed phenotypic abnormalities affecting their quality of life. Transgenic mice overexpressing Cre show no signs of phenotypic abnormality. Crosses of Plin and Cre models or injection of viruses to express or silence PCSK9 are expected to result in models with a mild phenotype. Individual experimental procedures described in this protocol are categorized as mild or moderate. The animal proposal is rated “Moderate”, although the majority of animals are anticipated to be exposed to mild distress.
III. Expected benefits
The obtained results will increase our understanding of the perilipin proteins and their role for LD accumulation in development of atherosclerosis. The murine Plin genes share high sequence homology with humans. Experimental results obtained in mice will therefore in large be representative for both species. Increased understanding of LD might reveal new molecular mechanisms that can be targeted with drugs to treat lipid diseases such as atherosclerosis.
IV. Number and type of animals
In total it is applied for 400 mice (mus musculus) of both genders and with variable age.
The mice will be genetically modified (Plin1-5, Ldlr, and ApoE knock-outs and transgenic with tissue-specific Cre expression).
V. Adherence to the 3Rs
Supplemental experiments will be performed in cell culture whenever possible; but whole animals are needed to understand the physiological role of the various Plin proteins. The laboratory mouse is a good model to study gene functions and human disease; with a physiology similar to humans and well-established methods for genetic modifications. Mice will be used for collection of organs, tissues and cells; sampling that would not be possible with human research subjects. Alternatives to animal use therefore do not exist. All lines are back-crossed to congenic strains (>N10) to reduce experimental variation and lower the number of animals needed for each experiment.
Cells in humans synthesize and store intracellular lipid droplets (LDs). LDs have multiple biological roles, e.g. as efficient reservoirs of energy and substrates for membrane and steroid hormone synthesis. Perilipin1-5 proteins are abundant on the LD surface and are important for regulation of LD degradation. Development of Atherosclerosis starts with accumulation of esterfied cholesterol, free cholesterol and cholesterol crystals in the arteria intima. The purpose of this project is to understand the biological role of LDs and Plin proteins in development of atherosclerosis using mice with deleted Plin1-5 genes.
According to §10, the purpose is: a) Basal research.
II. Expected distress
Mice with single mutation of Plin genes give offspring with normal Mendalian distribution with no observed phenotypic abnormalities affecting their quality of life. Transgenic mice overexpressing Cre show no signs of phenotypic abnormality. Crosses of Plin and Cre models or injection of viruses to express or silence PCSK9 are expected to result in models with a mild phenotype. Individual experimental procedures described in this protocol are categorized as mild or moderate. The animal proposal is rated “Moderate”, although the majority of animals are anticipated to be exposed to mild distress.
III. Expected benefits
The obtained results will increase our understanding of the perilipin proteins and their role for LD accumulation in development of atherosclerosis. The murine Plin genes share high sequence homology with humans. Experimental results obtained in mice will therefore in large be representative for both species. Increased understanding of LD might reveal new molecular mechanisms that can be targeted with drugs to treat lipid diseases such as atherosclerosis.
IV. Number and type of animals
In total it is applied for 400 mice (mus musculus) of both genders and with variable age.
The mice will be genetically modified (Plin1-5, Ldlr, and ApoE knock-outs and transgenic with tissue-specific Cre expression).
V. Adherence to the 3Rs
Supplemental experiments will be performed in cell culture whenever possible; but whole animals are needed to understand the physiological role of the various Plin proteins. The laboratory mouse is a good model to study gene functions and human disease; with a physiology similar to humans and well-established methods for genetic modifications. Mice will be used for collection of organs, tissues and cells; sampling that would not be possible with human research subjects. Alternatives to animal use therefore do not exist. All lines are back-crossed to congenic strains (>N10) to reduce experimental variation and lower the number of animals needed for each experiment.